Anti-Calcium Pump pan PMCA ATPase antibody [5F10]

• Supplier: Abcam
• Catalog: ab2825
• Prices: $403.00
• Sizes: 100 µl
• Host: Mouse
• Clonality: Monoclonal
• Isotype: IgG2a
• Clone: 5F10
• Applications: ICC/IF ICC/IF WB ICC/IF IHC-P FC B/N ELISA IHC-F IP
• Species Reactivities: Mouse, Rat, Sheep, Rabbit, Chicken, Hamster, Bovine, Cat, Dog, Human, Amphibians, Hamster, Primate
• Antigen: Full length native protein (purified) corresponding to Human Calcium Pump pan PMCA ATPase
• Description: Mouse Monoclonal
• Gene: ATP2B1
• Conjugate: Unconjugated

Product images

Immunocytochemistry/Immunofluorescence analysis of Calcium Pump pan PMCA ATPase shows staining in C6 cells. Calcium Pump pan PMCA ATPase staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or ab2825 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

Immunocytochemistry/Immunofluorescence analysis of Calcium Pump pan PMCA ATPase shows staining in HeLa cells. Calcium Pump pan PMCA ATPase staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or ab2825 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

Immunocytochemistry/Immunofluorescence analysis of Calcium Pump pan PMCA ATPase shows staining in U251 cells. Calcium Pump pan PMCA ATPase staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or ab2825 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

ab2825 Immunoprecipitate Calcium Pump pan PMCA ATPase in human whole cell lysate. 1,000,000 cells were lysed and incubated with primary antibody at 4µg/mg lysate and Protein A matrix for 20 hours at 4°C. For western blotting an undiluted Abcam`s ab1162, Rabbit polyclonal to DDDDK tag was used.See Abreview

Overlay histogram showing Jurkat cells stained with ab2825 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2825, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used with ab2825 (1/20 dilution) under the same conditions.

ab2825 at a 1/200 dilution detecting Calcium Pump pan PMCA ATPase in human monocytes by Flow Cytometry. The secondary used was an Alexa-Fluor 488 conjugated goat anti-mouse IgG (H+L) used at a 1/500 dilution.See Abreview

ab2825 staining Calcium Pump pan PMCA ATPase in human monocytes by Immunocytochemistry/ Immunofluorescence.Cells were fixed, permeabilized, blocked with 2% BSA for 30 minutes at 25°C and then incubated with ab2825 at a 1/250 dilution for 1 hour at 25°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-mouse IgG (H+L) used at a 1/1000 dilution.See Abreview

ab2825 staining Calcium Pump pan PMCA ATPase (green) in Mouse RAW 264.7 cells by ICC/IF (Immunocytochemistry/ Immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized in 0.1% Triton X-100 in 2% BSA for 15 minutes and blocked with 2% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/200 in PBS + 2% BSA) for 18 hours at 4°C. An Alexa Fluor®488-conjugated Chicken anti-rabbit IgG polyclonal (H&L) (1/750) was used as the secondary antibody. WGA (red) = Wheat Germ Agglutinin See Abreview

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human brain tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing PMCA ATPase ab2825 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing PMCA ATPase ab2825 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing PMCA ATPase ab2825 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Product References

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